Huang L C Lora, Chang Huan-Cheng
Institute of Atomic and Molecular Sciences, Academia Sinica, P.O. Box 23-166, Taipei, Taiwan 106, Republic of China.
Langmuir. 2004 Jul 6;20(14):5879-84.
Methods have been developed to immobilize proteins onto the surfaces ofnanodiamonds with an average size of 5 +/- 1 nm. The immobilization started with carboxylation/oxidization of diamonds with strong acids, followed by coating the surfaces with poly-L-lysine (PL) for covalent attachment of proteins using heterobifunctional cross-linkers. The feasibility of this approach is proven with fluorescent labeling of the PL-coated diamonds by Alexa Fluor 488 and subsequent detection of the emission using a confocal fluorescence microscope. Immobilization of proteins onto the surfaces is also demonstrated with yeast cytochrome c, which possesses a free SH group for linkage and a characteristic Soret absorption band for observation.
已开发出将蛋白质固定到平均尺寸为5±1纳米的纳米金刚石表面的方法。固定过程首先用强酸对金刚石进行羧化/氧化，然后用聚-L-赖氨酸（PL）包覆表面，以便使用异双功能交联剂共价连接蛋白质。通过用Alexa Fluor 488对PL包覆的金刚石进行荧光标记并随后使用共聚焦荧光显微镜检测发射光，证明了该方法的可行性。还用酵母细胞色素c证明了蛋白质在表面的固定，酵母细胞色素c具有用于连接的游离SH基团和用于观察的特征性Soret吸收带。
